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. 1977 Feb;4(2):339–352. doi: 10.1093/nar/4.2.339

Sequence analysis of T1 ribonuclease fragments of 18S ribosomal RNA by 5'-terminal labeling, partial digestion, and homochromatography fingerprinting.

M Fuke, H Busch
PMCID: PMC342436  PMID: 190590

Abstract

The method employed to determine the sequence of a T1 RNase fragment, A-A-A-A-A-U-A-A-C-A-A-U-A-C-A-Gp, from Novikoff rat hepatoma 18S ribosomal RNA is described. This method is applicable to any oligoribonucleotide produced by specific endonucleases that leave the newly cleaved 5'-end free for labeling with polynucleotide kinase and gamma-(32p)-ATP. The (32p)-labeled oligoribonucleotide is subjected to partial endonucleolytic digestion and fractionated by two-dimensional homochromatography fingerprinting. The nucleotide sequence is determined by following mobility shifts of the labeled and partially digested oligoribonucleotides in homochromatography fingerprinting.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arima T., Uchida T., Egami F. Studies on extracellular ribonucleases of Ustilago sphaerogena. Characterization of substrate specificity with special reference to purine-specific ribonucleases. Biochem J. 1968 Feb;106(3):609–613. doi: 10.1042/bj1060609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Egami F., Takahashi K., Uchida T. Ribonucleases in taka-diastase: properties, chemical nature, and applications. Prog Nucleic Acid Res Mol Biol. 1964;3:59–101. doi: 10.1016/s0079-6603(08)60739-4. [DOI] [PubMed] [Google Scholar]
  3. Eladari M. E., Galibert F. Sequence determination of 5'-terminal and 3'-terminal T1 oligonucleotides of 18-S ribosomal RNA of a mouse cell line (L 5178 Y). Eur J Biochem. 1975 Jun 16;55(1):247–255. doi: 10.1111/j.1432-1033.1975.tb02157.x. [DOI] [PubMed] [Google Scholar]
  4. Fuke M. Allocation of 15 RNase T1-resistant large oligonucleotides of MS2 RNA. J Biochem. 1976 Apr;79(4):731–737. doi: 10.1093/oxfordjournals.jbchem.a131125. [DOI] [PubMed] [Google Scholar]
  5. Fuke M., Busch H. A T1 ribonuclease fragment present in 18 S ribosomal RNA of Novikoff rat ascites hepatoma cells and absent from 18 S ribosomal RNA of HeLa cells. J Mol Biol. 1975 Dec 5;99(2):277–281. doi: 10.1016/s0022-2836(75)80145-8. [DOI] [PubMed] [Google Scholar]
  6. Fuke M., Busch H., Rao P. N. Evolutionary trends in 18S ribosomal RNA nucleotide sequences of rat, mouse, hamster and man. Nucleic Acids Res. 1976 Nov;3(11):2939–2957. doi: 10.1093/nar/3.11.2939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fuke M. Introduction of specific cleavages into RNAs of RNA bacteriophages for determination of base sequences. Proc Natl Acad Sci U S A. 1974 Mar;71(3):742–745. doi: 10.1073/pnas.71.3.742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Khan M. S., Maden B. E. Nucleotide sequences within the ribosomal ribonucleic acids of HeLa cells, Xenopus laevis and chick embryo fibroblasts. J Mol Biol. 1976 Feb 25;101(2):235–254. doi: 10.1016/0022-2836(76)90375-2. [DOI] [PubMed] [Google Scholar]
  9. Ling V. Fractionation and sequences of the large pyrimidine oligonucleotides from bacteriophage fd DNA. J Mol Biol. 1972 Feb 28;64(1):87–102. doi: 10.1016/0022-2836(72)90322-1. [DOI] [PubMed] [Google Scholar]
  10. MARKHAM R., SMITH J. D. The structure of ribonucleic acid. I. Cyclic nucleotides produced by ribonuclease and by alkaline hydrolysis. Biochem J. 1952 Dec;52(4):552–557. doi: 10.1042/bj0520552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Maniatis T., Jeffrey A., Kleid D. G. Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1184–1188. doi: 10.1073/pnas.72.3.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Richardson C. C. Phosphorylation of nucleic acid by an enzyme from T4 bacteriophage-infected Escherichia coli. Proc Natl Acad Sci U S A. 1965 Jul;54(1):158–165. doi: 10.1073/pnas.54.1.158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Székely M., Sanger F. Use of polynucleotide kinase in fingerprinting non-radioactive nucleic acids. J Mol Biol. 1969 Aug 14;43(3):607–617. doi: 10.1016/0022-2836(69)90362-3. [DOI] [PubMed] [Google Scholar]
  14. Takanami M. Analysis of the 5'-terminal nucleotide sequences of ribonucleic acids 1. the 5'-termini of Excherichia coli ribosomal RNA. J Mol Biol. 1967 Jan 28;23(2):135–148. doi: 10.1016/s0022-2836(67)80022-6. [DOI] [PubMed] [Google Scholar]

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